Delta&#39;-3, 11, 20-triketo-17-hydroxy-21-oxygenated steroids of the pregnane series and processes of preparing them



ite StamsPawh O h A'-3,11,20 TRIKETO-17-HYDROXY 21 OXYGEN- ATED STEROIDS OF THE PREGNANE SERIES AND PROCESSES F PREPARING THEM Evelyn H. Wilson, Highland Park, and Max Tishler,

Westfield, NJ., assignors to Merck & Co., Inc., Rahway, N..l., a corporation of New Jersey No Drawing. Application January 14, 1355 Serial No. 481,365

The terminal portion of the term or the patent suhse quent to December 4, 1973, has been disclaimed 4 Claims. (Cl. zen-491.45

This invention is concerned generally with A -unsaturated compounds of the pregnane series and with proc-.

asses of preparing them. More particularly, it relates to 3,11-diketo-steroids of the pregnane series having a 'double bond connecting the 6-1 and C-2 carbon atoms, and which may also have a M-unsaturated linkage, and with processes of preparing these novel n -steroid compoundsv These new A -3,11-diketo-steroids have been demonstrated to possess pharmacological activity similar to that shown by cortisone and are thus of value in the treatment of conditions which heretofore responded to the administration of the adrenal hormone, cortisone. Moreover, in addition to possessing cortisone-activity, these new A -3, ll-diketo-steroids difier from cortisone in not possessing any appreciable sodium or water retention action. Accordingly, these A -3,l1-diketo-steroid compounds are especially efiective in the treatment of arthritis and related diseases since they can be administered for their cortisone action without producing undesired metabolic efiects such as edema, which results from the sodiumand water retention action of cortisone. t

This application is-a continuation-in-part of our copending applications Serial No. 299,902, filed July 19, 1952, and Serial No. 227,102, filed May 18, 1951, now US. Patent No. 2,773,075.

These A -3,11-diketo-steroid compounds may be chem ically represented, insofar as rings A, B and C are con-' 7 corned, as follows:

The n -3,1l-diketo-steroid compounds, subject of the present invention, can be prepared by reacting the corresponding 2-halo-3,1l-diketo-steroid compound with a dehydrohalogenating agent. This reaction may be chemically represented, insofar as the changes taking place in rings A, B and C are concerned, as follows:

wherein X is halogen.

The 2-halo3,11-diketo-steroid compounds used as'starting materials include 2-halo-3,11,20-triketo-17-hydroxy- 21-acyloxy-steroid compounds as for example 2-bromo- .3,11,20 -triketo-17a-hydroxy 21 acyloxy-allopregnanes such as 2-bromo-3,1l,ZO-triketo-l7a-hydroxy-2l-acetoxyallopregnane, 2 bromo-3,11,20-triketo-17a-hydroxy 21- 2 propionoxy-allopregnane, 2 brorno 3,l1,20-triketo17 ahydroxy-Zl-benzoxy-allopregnane, 2-halo- 3,11,20-triketo- 17 whydroxy-Zl-acyloXy-pregnanes, such as 2-bromo-3,11, ZO-triketo-l7a-hydroxy 2l-acetoxy-pregnane, 2 ammo-3,. 11,20-triketo-l7a-hydroxy 21 propionoXy-pregnane, 2- bromo-3,1 1,20-triketo 17a hydroxy 21 benzoxy-preg' nane, 2,4 dibromo 3,11,20 triketo 17a hydroxy 21- acyloxy-allopregnanes, such as 2,4-dibromo-3,l1,20-t1iketo-l7a-hydroxy-2l-acetoxy-allopregnanes, 2,4-dibromo- 3,11,20-triketo 17a hydroxy 7 21 propionoXy-allopregnane, 2,4-dibromo-3,11,20-triketo-17a-hydroxy-2l-benz oxy-allopregnane, -2,4'-' dibromo 3,11,20 triketo-lh-hydroxy-Zl-acyloxy-pegnanes, such as 2,4-dibromo-3,i-1{ 20-triketo-17a-hydroxy 7 21 acetoxy-pregnane, 2,4-dibromo-3,l1,20-triketo-17a hydroxy-2l-propionoxy pregnane, 2,4-dibromo-3,11,20-t1iketo-17a-hydroxy benzoxypregnane, as well as the corresponding chloro and iodo derivatives and the like.

The reaction between the 2-halo-3,1i-diketo-steroid compound and the dehydrohalogenating agent can be conveniently conducted by reacting the 2-halo-3,I1-diketosteroid compound with a tertiary amine such as pyridine, collidine, and the like. It is preferred, however, to utilize 2,4-dinitrophenylhydrazine or semi-carbazide in conjunction with pyruvic acid as the dehydro-halogenating agent. When the 2-halo-3,l1,20-triketo-l7-hydroxy-21 acyloxy-steroid is a 2,2-dihalo-3,11,20-triketo-17-hydroxy; Zl-acyloxy-steroid of the pregnane series, such as 2,24 tribromo 3,11,20 triketo 17a hydroxy 21 acetoxy-. allopregnane, an alkali metal iodide, such as sodium iodide, may be used as the dehydrohalogenating ageut. When the 2,4-dinitrophenylhydrazine is used, the Z-halo;

3,11-diketo-steroid compound is preferably brought into contact with the 2,4-dinitrophenylhydrazine in a medium comprising glacial acetic acid, at a temperature of about 5055 C. The intermediate 2,4-dinitrophenylhydrazone of A -3,11,20-triketo-17-hydroxy-2l-acyloxy-steroid compound is conveniently recovered from the glacial acetic acid reaction mixture by diluting the latter with water and cooling, whereupon the hydrazone compound crys-t tallizes as an orange solid, which is recovered by filtration and dried. The crude hydrazone, thus obtained is then heated with a solution of pyruvic acid in acetic acid at an elevated temperature of approximately -85? 'C. At this temperature, the hydrolysis of the hydrazone is ordinarily complete in less than one hour, but the hy drolysis reaction is preferably carried out for a period of about five to six hours. 'Ihe'product is conveniently recovered from the hydrolysis-solution by evaporating the latter to small volume and diluting the residual solu tion with chloroform. The by-product impurities which precipitate are separated from the resulting solution by filtration, and the chloroform filtrate is then washed with water and aqueous alkaline solution to remove additional impurities. This chloroform solution can then be evaporated, if desired, to obtain the corresponding A -3,1l,20- triketo-l7,21-dihydroxy-steroid compound such as N- 3,1 1,20-triketo-17a,2l-dihydroxy-allopregnene, A -3, 11,20 triketo :,21 dihydroxy-pregnene, A -3,11,20-triketo- 1711,2l-clihydroxy-pregnadiene, etc. It is ordinarily preierred, however, to treat the chloroform solution an acylating agent, as, for example, benzoic anhydride, a lower alkanoic anhydride, such as acetic anhydride and pyridine, a lower alkanoyl halide, benzoyl halide, and the like, thereby acylating the 2l-hyd1'oxy substituent. The acylated product is readily separated from the chloroform solution by conventional means, and purified by recrystallization from a solvent such as ethyl acetate .to give the A -3,11-diketo-steroid compound as, for example, A 3,11,20 triketo 17a hydroxy 21 acylory-allopregnenes, such as A -3,'11,ZO-triketo-17a-hydrony-21- Patented Ap'r.'19,'196 0i 2,983,515 a J. V

eetexy-atlc re ae; 1-3,: ao-inkedr'la-h dmx -zipropi noxy-allopregnene, 4 -3, 1 l ,ZO-triketo-l 7e-hydroxy: ZI-benzoxy-allopregnene, A -3,l l,20-triketol7a-hydroxy- 2l-acyloxy-pregnenes, ti -3,11,20-triketo-l7a-hydroxy-2lacetoxy-pregnene, Is -3,1 1,20-triketo-17a-hydroxy-2 l-propionoxy-pregnene, A -3,ll,20 trilteto 17a hydroxy=2lbenzoxy-pregnene, d -3,1 1,20-triketo 17a hydroxy-Z aeyloxy-pregnadienes, such as A -3,ll,20-triketo-l7ahydroxy-Zl-alkanoyloxy pregnadienes, A -3,ll,20 trilgeto-l7e-hydroxy-21-acetoxy pregnadiene, N -3,11,20- 20-triketo-17a-hydroxy-2l-butyroxy-pregnadiene, d -3,- 11,20 triketo-l7a-hydroxy-2l-benzoxy-pregnadiene, and the like.

The following examples illustrate methods of carrying out the present invention, but it is to be understood that these examples are given for purposes ofillustration and not of limitation.

e ample .A. mix r o 4- s- 1 2-b o no-3 l .20: triketo-17cz-hydroxy-21-acetoxyallopregnane, 2.2 g." of 2,4-dinitrophenylhydraziue and 125 cc. of glacial acetic acid is heated, with agitation under a nitrogen atmosphere for a period ofapproximately three hours at a temperature of approximately 50-55 C. Three hundred and seventy-five cubic centimeters of distilled water is added to the reaction mixture, and the resulting mixture is cooled and maintained at a temperature of about 0 C. for a period of approximately two hours. The orange solid which precipitates is recovered by filtration, washed well with water and air dried, to give the crude 2,4-dinitrophenylhydrazone of l -3,11,20-triketo-17a-hydroxy- 21-acetoxy-allopreg'nene.

This crude hydrazone is mixed with 175 cc. aqueous pyruvic acid and cc. of glacial acetic acid, and the mixture is heated under nitrogen, with agitation,

' at a temperature of about 80-85 C. for a period of #PPIOXimately five and one-half hours.

(A clear solution is obtained after heating the mixture for aperiod of approximately forty-five minutes.) The reaction solution is evaporated in vacuo to a small volume, and the residual solution is diluted with a large quantity of chloroform. The solid material which precipitates is removed by filtration, and the chloroform filtrate is extracted with two portions of water, with three portions of dilute aqueous potassium bicarbonate solution, and finally with another portion of water. Evaporation of this washed chloroform solution to dryness gives A-3,11,20-t1'lkt0r 170:,2l-dihydroxy-allopregnene. Alternatively, the washed chloroform solutionis evaporated to onerhialf its initial volume at aPPSoximately atmospheric pressure.

' To the dark chloroform solution thus obtained are added 20 cc. of acetic anhydride and 5 cc. of pyridine.

. The resulting solution is maintained at approximately room temperature for a period of about fifteen hours. The reaction solution is then shaken with water, and the chloroform layer is washed successively with a 2.5 N aqueous solution of hydrochloric acid, with water, with dilute aqueous potassium bicarbonate solution, and finally with water. The washed chloroform solution is dried over anhydrous calcium sulfate (Drierite), and the dry chloroform solution is then passed through a column containing 100 g. of acid-washed alumina. The chloroforrn eluates from the initial solution and washings are combined vandtreated with activated charcoal (Norit),

f ltered and the filtered solutionis evaporated to dryness in vacuo. "The residual material is recrystallized from' ethyl acetate to give 900 mg. of A -3,ll,20-triketo-l7ahydroxy-Zl-acetoxy-allopregnene which is obtaiued'in the form of fine white needles; M.P. 253-256 C. (dec jcl -=-!-ll5" (0.2% in chloroform); C Hg-OI-I 22.70. A.; Log E. 4.06. Analysis: Calcd for C l-1 50 C, 68.63; H, 7.51. Found: C, 68.53; H, 7.66.: p r' 2 bromo-3,l.1,201triketo- 17a hydroxy 2'l-acetoxyallopregnane used as starting material in this example may be prepared as follows: six-hundred milligrams of palladium oxide and 12 cc. of 0.001 N aqueous solution of potassium hydroxide are added to a suspension of 7.25 g. (0.018 mole of At-3,1l,20-triketo-17e-hydroxy- 2l-acetoxy-pregnene (cortisone acetate) in 600 cc. of methanol. The mixture is then reacted with hydrogen at a pressure of approximately 40 pounds per square inch While maintaining the temperature of the reactants at approximately room temperature. The absorption of ceases after approximately one-half hours; the amount of hydrogen absorbed corresponds to 1 mole.- The reaction mixture is filtered, thereby removing the catalyst, and the catalyst is Washed thoroughly with chloroform; A few drops of ethanolic hydrogen chloride are added to the combined filtrate and washings, and the resulting solution is evaporated to dryness in vacuo. The residual material is dissolved in chloroform, and the solutionis again evaporated to dryness. The residnai material is then recrystallized twice from acetone to give substantially pure 3,l1,20triketo-17e-hydroxy-2l-aeetoxymaterial is triturated with low-boiling petroleum ether,

and the insoluble material is recovered by filtration and recrystallized from ethyl acetate to give 2-bromo-3,11,20 triketo-17m-hydroxy-2l-acetoxy-allopregnane.

Example 2 rial is recrystallized from ethyl .acetateto'give substantially pure A -3,11,20-triketo- 17a hydroxy-21-acetoxypregnene; M.P.'244246 C.

The 2-bromo-3,1l,ZO-triketo-l7a-hydroxy-2l-acetoxy- V pregnane, used as starting material in this example, may.

be prepared as follows: a solution containing 39.6 g. of

bromine in 300 cc. of acetic acid is added to a solution containing g. of 3,l1,20-triketo-17a-acetoxy-pregnane dissolved in 1500 cc. of acetic acid. When the reaction is substantially complete, the solution is immediately poured into water, and the resulting suspension is extracted with chloroform. The chloroform extract is washed with water, evaporated'to dryness, and the residual material crystallized from acetone-ether to give as a substantially pure crystalline product the 4-bromo-3,l1,20- triket'o-17u-hydroxy-2l-acetoxy pregnane.

The mother liquor from this crystallization procedure is dissolved in benzene and chromatographed over acidwashed alumina, and the chromatogram eluted with mixtures of ether and chloroform. The eluate is evaporated to dryness, and the residual crystalline material is recrystallized from ethyl acetate to give -2 -bro mo-3,1l,20-' triketo-l7e-hydroxy-2l acetoxy pregnane.

Example" 3 mixture is extracted three times with chloroform,- and the combined chloroform extracts are dried and the chloroform evaporated therefrom in vacuo. The residual material is dissolved in benzene and chromatographed over 15 g. of acid-washed alumina. The chromatogram is eluted with mixtures of ether and chloroform, and the combined eluates are evaporated to dryness. The residual crystalline material is recrystallized from ethyl acetate to give substantially pure A -3,11,20-triketo-lh-hydroxy- 21-acetoxy-pregnadiene; M.P. 226-228 C. @One hundred milligrams of A -3,11,20-triket-17ahydroxy-2l-acetoxy-pregnadiene is dissolved in a mixture of 1.0 cc. of benzene and 1.0 cc. of 1.1 N methanolic potassium hydroxide, and the solution is allowed to stand at room temperature for a period of about 10 minutes. Thesolution is then acidified with acetic acid, the henzene is "evaporated in vacuo, and'the' residual material is recrystallized from ethyl acetate'to give substantially pure A -3,11,20-triketo-17a,2l-dihydroxy-pregnadiene; M.P. 207-214 C. 1

The A -4-bromo-3,11,20-triketo-17a-hydroxy-2l-acetoxy-pregnene, used as starting material in this example, may be prepared as follows: four hundred mg. of A 3,11,20-triketo-17a-hydroxy-2l-acetoxy-pregnene is dissolved in 50 cc. of glacial acetic acid containing three drops of 30% hydrobromic acid in glacial aceticacid, and to the stirred solution is added a solution containing 0.61 ml. bromine (190 mg.) in 5 ml. of glacial acetic acid over a ten minute period. Five minutes after the addition of bromine is completed, the reaction mixture is poured into 400 ml. of ice water, and the aqueous mixture is extracted three times with chloroform. The chloroform extracts are combined, washed with aqueous sodium bicarbonate solution, then with water, dried and evaporated to dryness in vacuo to give A -4-bromo-3,11,20- triketo-17a-hydroxy-2l-acetoxy-pregnene.

Example 4 202 mg. of 3,11,20-triketo-17a-hydroxy-2l-acetoxyallopregnane, prepared as described in Example 1 hereinabove, is dissolved in 15 cc. of chloroform, and to the resulting mixture is added dropwise 4.3 cc. of a 0.0011 N chloroform solution of bromine in cc. of chloroform. This solution is allowed to stand at room temperature for about fifteen hours, and the chloroform solution evaporated to dryness in vacuo to give 2,4-dibromo- 3,11,20-triketo-17e-hydroxy-2l-acetoxy-allopregnane.

This 2,4-dibromo compound is dissolved, without purification in 25 cc. of glacial acetic acid and to the solution is added 282 mg. of 2,4-dinitrophenylhydrazine. The resulting solution is heated under'nitrogen at 50-55 C. for two hours, and the reaction solution is worked up in the conventional manner to give the desired A -3,11, 20-triketo-l7a-hydroxy-2l-acetoxy-pregnadiene 3-(2,4-dinitrophenylhydrazone). This product is hydrolyzed in accordance with the procedure set forth in Example 1 hereinabove to give la -3,11,20-triketo-l7a,21dihydroxypregnadiene.

Example 5 1.213 g. of 3,11,20-triketo-17a-hydroxy-2l-acetoxyallopregnane (0.003 mole) is dissolved in cc. of chloroform, 2 drops of 1.3 N hydrogen bromide in acetic acid is added, and to the solution is added, dropwise with stirring, 0.009 mole of bromine in 25 cc. of chloroform At the completion of the bromination, 750 mg. anhydrous sodium acetate is added to the reaction mixture and the resulting mixture is stirred for one hour'at room temperature. The reaction mixture is filtered and the filtrate is evaporated to dryness in vacuo. The residual material, c omprising 2,2,4-tribromo-allodihydro cortisone acetate, is dissolved in 250 cc. reagent acetone containing 5 g. of sodium iodide. The resulting mixture is heated under reflux for a period of 24 hours, an additional 5 g. of sodium iodide is added to the-reaction mixture, and the resulting mixture is heated under reflux for an additional 16 hours. The reaction mixture is evaporated to dryness in vacuo, and the residual material is partitioned between water and chloroform. The chloroform layer is washed with aqueous sodium thiosulfate solution, aqueous sodium bicarbonate solution, and Water, and is dried by distillation. The residual material is heated under reflux for a period of one-half hour with 35 cc. of gamma collidine. The reaction solution is poured into dilute aqueous sulfuric acid solution, and the aqueous mixture is extracted with chloroform. The chloroform extracts are washed with water, dried, and the residual material is treated with 3 cc. of acetic anhydriderand 0.5 cc. of pyridine for about 60 hours at ethyl acetate to give A -3,11,20+triketo-l7a-hydroxy-2lv acetoxy-pregnadiene; M.P. 210--2l8' C. This material may be purified further by chromatography if desired to give material melting at 226-22 8 C.

Example 6 530 mg. of 3,11,20-triketo-17a-hydroxy-21-acetoxyallopregnane (0.0013 mole) is-dissolved in 50 cc. of dry chloroform. The solution is stirred at room temperature while adding thereto, dropwise, 10 cc. of a solution of bromine in chloroform containing 0.0043 mole. The reaction solution thus obtained is evaporated to dryness in vacuo, and the residual material is dissolved in 50 cc. of acetonitrile. To this solution is added 1.0 g. of semicarbazide free base (0.013 mole) and 2 cc. of glacial acetic acid. The resulting mixture is stirred at room temperature for 48 hours, and then heated under reflux for seven hours. The reaction mixture is evaporated to dryness in vacuo,.and the residual material is slurried with water, filtered and washed with water. The solid material thus obtained is dissolved in 28 cc. of glacial acetic acid, this solution is diluted with 12 cc. of water and 1 cc. of pyruvic acid is added to the solution. The resulting mixture is maintained at room temperature for about 60 hours, and is then evaporated to dryness under reduced pressure. The residual material is slurried with chloroform, the insoluble material is removed by filtration, and the filtered chloroform solution is washed with water, dilute aqueous sodium bicarbonate solution, then with water and dried. The chloroform is evaporated in vacuo to give a crude mixtxure comprising A -3,11,20-lllkl0-17uhydroxy-ZI-acetQXy-pregnene and A -3,11,20-triketo-l7uhydroxy-Zl-acetoxy-pregnadiene. This mixture is recrystallized once from acetone and once from methanol to give, as the major product A -3,11,20-triketo-17a-hydroxy- 21-acetoxy-pregnene; M.P. 237240 C. dec. The residual mother liquor containing the A -3,11,20-triketo-17uhydroxy-2l-acetoxy-pregnadiene may be purified by chromatography. to obtain the latter compound in substantially pure form.

Example 7 1.011 g. (.0025 m.) of allodihydro cortisone acetate is dissolved in 35 cc. freshly distilled chloroform and treated all at once at noom temperature with .00825 mole of bromine in 15 cc. chloroform. Decolorization is completed in about 10 minutes. The chloroform solution is washed with water, potassium bicarbonate solution, water, and finally with brine. The yellow solid that precipitates is filtered, washed well with water, and dried to give a product comprising a major proportion of A -2-iodo-3,l1,ZO-triketo-17a-hydroxy-2l-acetoxy-pregnene and a minor proportion of A -3,11,20-triketo-17ahydroxy-Zl-acetoxy-pregnadiene.

Example 8 7 prepared by reacting allodihydrqcgrtisone acetate with semicarliazide in accordance with known methods, is dissolvedin25cc. t glacial acetic acid. This solution is" maiutainatroom temperature while adding thereto;

dropwise with stirring, cc. of a solution of bromine in glacial acetic acid containing 0.002 mole. One drop of 1 N hydrogen bromide in acetic acid is added to the bromination mixture, and the resulting mixture is stirred at room temperature until decolorization is complete, about 10 minutes being required. Nitrogen is then bubbled through the decolorized mixture for minutes, and, while continuing to bubble nitrogen, the mixture is warmed at -50 C. for four hours. Dilution of the reaction mixture with water and filtration of the resulting suspension gives a mixture of semicarbazones' com; prising A d-3,1l,ZO-triketo-l'ld-hydtoxy-Zl-acetoxy-preg nadieue 3-semicarbarzone. This productis hydrolyzed in accordance with the procedure 'set forth in Example 1 2. 3,11,ZO-trilretgd7a-hydi oxy-steroids of the pregnane series having adouhle hondconneeting' the and'G-Z carbon atonis,' and 'a 6-21 substituent selected from the group which consists of hydroxynnd lower hydrocarbon carbonyloxy radicals. i i

3. Ring A unsaturated 3,11,20-t1ik6t0-17oz-hYdl'OXY- steroids of the allo'pr egnaue series having a-double bond connecting the C.1 and C-2 carbon atoms, and a lower alkanoyloxy substituent at 0-21. I

4. Ring 'A unsaturated 3,11,20-triketo-17a-hydroxysteroids of the ail'opregnane series having a double bond connecting the 0-1 and GT2 carbon atoms,'and a hydroxy substituent at 6 -21.

Regen-cranes (fJited in the file of this patent UNI??? TTE PATENT TF7. -:-t: 2.590.2 8 s ri e e' 'Ap 1. 195 2 703 805 Kesen sreaz. M .1 5 2,768,189 Nomine V o r. 2 3, 1955 2,773 t) Wilson et al. n. Dec. 4, 1956 2,774,775 Korman et a1. Dec. 18 1956 2.852539 Qay' et'al, Sept. l6 1958' T ERL E ER Djerassi; Jour. Chem. Soc. 69, 2404-11) 1947 Rosenlgrang: Jour. Chem. Soc. 72, 1046 (1950). Rosenlgranz: Jour. Chem. Soc. 72, 4081- (1.950 r a 

1. THE PROCESS WHICH COMPRISES REACTING A 2-HALO-3-11DIKETO-STEROID OF THE PREGNANE SERIES WITH A DEHYDROHALOGENATING AGENT TO FORM THE CORRESPONDING $1-3,11DIKETO-STEROID.
 2. 3,11,30-TRIKETO-17-A-HYDROXY-STEROIDS OF THE PREGNANE SERIES HAVING A DOUBLE BONDF CONNECTING THE C-1 AND C-2 CARBON ATOMS, AND A C-21 SUBSTITUENT SELECTED FROM THE GROUP WHICH CONSISTS OF HYDROXY AND LOWER HYDROCARBON CARBONYLOXY RADICALS. 